Campbell-Thompson Martha L, Heiple Tiffany, Montgomery Emily, Zhang Li, Schneider Lynda
Department of Pathology, Immunology, and Laboratory Medicine, University of Florida, USA.
J Vis Exp. 2012 May 23(63):e4068. doi: 10.3791/4068.
Estimates of islet area and numbers and endocrine cell composition in the adult human pancreas vary from several hundred thousand to several million and beta mass ranges from 500 to 1500 mg. With this known heterogeneity, a standard processing and staining procedure was developed so that pancreatic regions were clearly defined and islets characterized using rigorous histopathology and immunolocalization examinations. Standardized procedures for processing human pancreas recovered from organ donors are described in part 1 of this series. The pancreas is processed into 3 main regions (head, body, tail) followed by transverse sections. Transverse sections from the pancreas head are further divided, as indicated based on size, and numbered alphabetically to denote subsections. This standardization allows for a complete cross sectional analysis of the head region including the uncinate region which contains islets composed primarily of pancreatic polypeptide cells to the tail region. The current report comprises part 2 of this series and describes the procedures used for serial sectioning and histopathological characterization of the pancreatic paraffin sections with an emphasis on islet endocrine cells, replication, and T-cell infiltrates. Pathology of pancreatic sections is intended to characterize both exocrine, ductular, and endocrine components. The exocrine compartment is evaluated for the presence of pancreatitis (active or chronic), atrophy, fibrosis, and fat, as well as the duct system, particularly in relationship to the presence of pancreatic intraductal neoplasia. Islets are evaluated for morphology, size, and density, endocrine cells, inflammation, fibrosis, amyloid, and the presence of replicating or apoptotic cells using H&E and IHC stains. The final component described in part 2 is the provision of the stained slides as digitized whole slide images. The digitized slides are organized by case and pancreas region in an online pathology database creating a virtual biobank. Access to this online collection is currently provided to over 200 clinicians and scientists involved in type 1 diabetes research. The online database provides a means for rapid and complete data sharing and for investigators to select blocks for paraffin or frozen serial sections.
成体人类胰腺中小岛的面积、数量及内分泌细胞组成的估计值从几十万到几百万不等，β细胞团质量范围为500至1500毫克。鉴于这种已知的异质性，我们开发了一种标准的处理和染色程序，以便通过严格的组织病理学和免疫定位检查清晰界定胰腺区域并对小岛进行特征描述。本系列第1部分描述了从器官捐献者处获取的人类胰腺的标准化处理程序。胰腺被处理成3个主要区域（头部、体部、尾部），然后进行横切。胰腺头部的横切根据大小进一步划分，并按字母顺序编号以表示子部分。这种标准化使得能够对头部区域进行完整的横断面分析，包括含有主要由胰多肽细胞组成的小岛的钩突区域至尾部区域。本报告是该系列的第2部分，描述了用于胰腺石蜡切片连续切片及组织病理学特征描述的程序，重点是小岛内分泌细胞、复制及T细胞浸润。胰腺切片的病理学旨在对腺泡、导管及内分泌成分进行特征描述。评估腺泡部分是否存在胰腺炎（急性或慢性）、萎缩、纤维化和脂肪，以及导管系统，特别是与胰腺导管内瘤变的关系。使用苏木精-伊红染色和免疫组化染色评估小岛的形态、大小和密度、内分泌细胞、炎症、纤维化、淀粉样蛋白以及复制或凋亡细胞的存在情况。第2部分描述的最后一个内容是提供染色玻片作为数字化全玻片图像。数字化玻片按病例和胰腺区域在一个在线病理学数据库中进行整理，创建一个虚拟生物样本库。目前，超过200名参与1型糖尿病研究的临床医生和科学家可以访问这个在线馆藏。该在线数据库提供了一种快速且完整的数据共享方式，供研究人员选择石蜡或冷冻连续切片的组织块。
